Mouse Anti-CCR5 Recombinant Antibody (clone RoAb13) (CAT#: PABL-345)

Recombinant Mouse Antibody (RoAb13) is capable of binding to The extracellular domain of CCR5. This antibody could block HIV infection, also blocks migration of monocytes in response to CCR5 binding chemokines and to inflammatory macrophage conditioned medium.


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Figure 1 The antibodies recognize a linear epitope in the N-terminal domain of CCR5.

Figure 1 The antibodies recognize a linear epitope in the N-terminal domain of CCR5.

Dilutions of the four monoclonal antibodies shown were tested for binding to hCCR51–31 coding for the N-terminal extracellular domain of CCR5 by ELISA. The left panel shows binding (as measured by OD at 405 nm) to plates coated with the N-terminal peptide coupled to avidin, while the right panel shows binding to avidin alone.

Chain, B., Arnold, J., Akthar, S., Brandt, M., Davis, D., Noursadeghi, M.,... & Govada, L. (2015). A linear epitope in the N-terminal domain of CCR5 and its interaction with antibody. PloS one, 10(6), e0128381.

Figure 2 R&D1801 and RoAb13 recognise overlapping core epitopes in the CCR5 N-terminal domain.

Figure 2 R&D1801 and RoAb13 recognise overlapping core epitopes in the CCR5 N-terminal domain.

A, R&D1801 or RoAb13 were incubated overnight with competing peptides 1–24 (1 μg/ml) in wells coated with hCCR5₁₋ ₃₁, and binding (shown as OD 405 nm) of antibody was then measured using a rabbit anti-mouse second layer. C—PBS used in place of competitive peptide. B, The peptide sequence of the 24 peptides tested in a. The blue bars shows all peptides which inhibited the binding by more than 50%. C, Diagrammatic representation of the core binding epitope recognized by the two antibodies, defined by peptides which inhibited binding to the whole N-terminal peptide by more than 50%.

Chain, B., Arnold, J., Akthar, S., Brandt, M., Davis, D., Noursadeghi, M.,... & Govada, L. (2015). A linear epitope in the N-terminal domain of CCR5 and its interaction with antibody. PloS one, 10(6), e0128381.

Figure 3 Binding of immune sera and RoAb13 to sulphated N-term CCR5.

Figure 3 Binding of immune sera and RoAb13 to sulphated N-term CCR5.

Dilutions of sera from immunized mice 1–5 or purified RoAb13 supernatant were tested by ELISA for binding to hCCR5₁₋ ₃₁, or an analogue of the same peptide modified by sulfation of tyrosines at position 10 and 14.

Chain, B., Arnold, J., Akthar, S., Brandt, M., Davis, D., Noursadeghi, M.,... & Govada, L. (2015). A linear epitope in the N-terminal domain of CCR5 and its interaction with antibody. PloS one, 10(6), e0128381.

Figure 4 Species specificity of RoAb13 and R&D1801.

Figure 4 Species specificity of RoAb13 and R&D1801.

A, The sequence of the N-terminal extracellular domain of CCR5 from human, rhesus Macaque or mouse CCR5. * identical amino acids—insertion in mouse sequence. B, Flow cytometry showing binding of R&D1801 or RoAb13 to CHO cells transfected with mouse (filled) or human (red line) CCR5. C, Flow cytometry showing binding of R&D1801 or RoAb13 or control to PBMC from rhesus macaques (indirect immunofluorescence, using Goat anti-mouse PE conjugate as second layer). The first panel shows binding of a commercially available PE-conjugated anti-rhesus CCR5 antibody (positive control).

Chain, B., Arnold, J., Akthar, S., Brandt, M., Davis, D., Noursadeghi, M.,... & Govada, L. (2015). A linear epitope in the N-terminal domain of CCR5 and its interaction with antibody. PloS one, 10(6), e0128381.

Figure 5 RoAb13 inhibits monocyte chemotaxis.

Figure 5 RoAb13 inhibits monocyte chemotaxis.

PBMC were incubated in the upper chamber of transwells, while the lower chamber contained medium alone, supernatant of unactivated macrophages, or supernatant of macrophages activated with LPS. RoAb13 Fab fragment or a control Fab fragement were added to some wells as shown. Migrating cells were collected from the lower chamber after three hours. Migration is shown as the proportion of cells in the PBMC premigration sample which migrate to the lower chamber during the assay.

Chain, B., Arnold, J., Akthar, S., Brandt, M., Davis, D., Noursadeghi, M.,... & Govada, L. (2015). A linear epitope in the N-terminal domain of CCR5 and its interaction with antibody. PloS one, 10(6), e0128381.

Figure 6 CCR5 MAb dose-response curves of MAb3952-resistant and wild-type Bal (A) and CC1/85 (B) HIV-1 strains in a PBMC-based assay.

Figure 6 CCR5 MAb dose-response curves of MAb3952-resistant and wild-type Bal (A) and CC1/85 (B) HIV-1 strains in a PBMC-based assay.

A total of 100,000 PBMC were infected with 800 pg p24 of the indicated virus strain in the presence of diluted CCR5 MAb MAb3952 (red), RoAb13 (blue), or 2D7 (black). After incubation for 6 days, virus production was determined using a p24 ELISA. NDC viruses are indicated by filled symbols and continuous lines, and MAb3952-resistant (MAb3952res) virus strains are indicated by open symbols and broken lines.

Jekle, A., Chhabra, M., Lochner, A., Meier, S., Chow, E., Brandt, M.,... & Heilek, G. (2010). Epitope switching as a novel escape mechanism of HIV to CCR5 monoclonal antibodies. Antimicrobial agents and chemotherapy, 54(2), 734-741.


Specifications

  • Immunogen
  • Human chemokine (C-C motif) receptor 5 (gene/pseudogene)
  • Host Species
  • Mouse
  • Derivation
  • Mouse
  • Type
  • Mouse IgG
  • Specificity
  • Human CCR5
  • Species Reactivity
  • Human
  • Clone
  • RoAb13
  • Applications
  • Block, ELISA, FC

Product Property

  • Purity
  • >95% as determined by analysis by SDS-PAGE
  • Concentration
  • Please refer to the vial label for the specific concentration.
  • Buffer
  • PBS
  • Preservative
  • No preservatives
  • Storage
  • Centrifuge briefly prior to opening vial. Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C long term. Avoid repeated freeze/thaw cycles.

Target

  • Alternative Names
  • CCR5; chemokine (C-C motif) receptor 5 (gene/pseudogene); CKR5; CCR-5; CD195; CKR-5; CCCKR5; CMKBR5; IDDM22; CC-CKR-5; C-C chemokine receptor type 5; chemr13; HIV-1 fusion coreceptor; chemokine receptor CCR5; C-C motif chemokine receptor 5 A159A

Product Notes

This is a product of Creative Biolabs' Hi-Affi™ recombinant antibody portfolio, which has several benefits including:

• Increased sensitivity
• Confirmed specificity
• High repeatability
• Excellent batch-to-batch consistency
• Sustainable supply
• Animal-free production

See more details about Hi-Affi™ recombinant antibody benefits.

Downloads

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Humanized Antibody

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TAB-10000S Anti-Human CCR5 Recombinant Antibody IF, IP, Neut, FuncS, ELISA, FC, ICC IgG4

Fc Glycosylation

CAT Product Name Application Type
Gly-106LC Recombinant Anti-Human CCR5 Antibody (Fc glycosylation) ELISA Mouse antibody

Neutralizing Antibody

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NEUT-267CQ Mouse Anti-CCR5 Recombinant Antibody (clone 45523.111) FC, IHC-P, Neut Mouse IgG2b
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For research use only. Not intended for any clinical use. No products from Creative Biolabs may be resold, modified for resale or used to manufacture commercial products without prior written approval from Creative Biolabs.

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